Wrap up of the purification and transformation protocol. Started the enzyme assays. One of which failed, consequently tried again.
Characterization results indicate weight of about 47kDa.
This is the figure from the successful enzyme assay.
This is the data results from the unsucessful enzyme assay.
Week 14 - 11/26/2012 Worked a bit on the surrogate procedure. Predominately spent time on the virtual screening of the target and creating a swiss model. Ligand binding continues to prove unsucessful. Will recheck gold coordinates next week.
Figure 1: Homology model achieved using the SWISS protocol.
Week 13 - 11/19/2012
112612 - Good - show some 'data' or image. Dr B Worked a bit on the latter portion of the surrogate protocol. No other work due to Thanksgiving Break. Data images pending.
Week 12 - 11/12/2012 Akhi, ok - would like to see graph or data from virtual here. -- Dr. B 11/19/12
Moved onto the surrogate protocol. Did the day one and day two steps.
mL required for 0.1 OD reading - 40mL.
(More specific data impending)
1.5 hours required to achieve .346 OD reading.
Weight of Bacteria - 1.5g , 1.19g, 1.16g.
Week 11 - 11/5/2012 Started the cloning and transformation protocol. Then decided it would be more efficient if Shane took care of most of it and I made more backup PCR products instead. First I though I found some leftover PCR sqared, so I tested it, however it was not PCR product at all (picture below confirms it) . Then I used PCR secondary and started the to create PCR squared. However, Shane tested it and found out that the PCR did not work.
Results indicate anonymous samples were not PCR squared. work.
Week 10 - 10/29/2012 PCR squared was finally a success. Two concentrations were achieved. PCR cleanup concentration 1: Gained through using 100uL of PCR product during cleanup.
PCR cleanup concentration 2: gained through using another 100uL of PCR product during cleanup.
Both concentrations are acceptable, though the 56.4 ng/uL is preferable. My group plans to utilize this latter.
Week 9 - 10/22/2012 Did two PCR cleanups. Unfortunately, both results yielded 0 ng as a concentration. This would mean that either I am missing some important component in my PCR protocols, or I am getting very unlucky. Will try another PCR cleanup next week.
Week 8 - 10/15/2012 Predominately dedicated time to Virtual Drug Screening Refresher. Results are posted on my Google docs. Was going to start PCR cleanup on Saturday but fell ill.
Week 7 - 10/8/2012 101612 - Akhi - ok good job. Good luck with PCR cleanup step and cloning. -- Dr. B
The final four lanes are my PCR^2. The results correlate with the results from PCR secondary and also match up with the predicted length of my gene sequence.
I also worked on virtual drug screening and finished part 1. Currently waiting for results.
Week 6 - 10/1/2012
100912 - Akhi, ok great. I am glad you guys got this to work. Crop your gels a little more so the gel is more visible. When you guys move on to cloning - use the NEW T4 DNA Poly and the new dGTP and new dCTP.
- Dr. B
Ran primary PCR and got these results.
PCR Primary Results
Lane 1 - 100bp ladder
Lane 2 - Shane's sample
Lane 3 - Mihir's sample
Lane 4 - My sample (not even visible), perhaps attributed to the low temperature I used during the pcr program.
Secondary PCR (done using Mihir's stock)
Lane 1 - 100bp ladder
Lane 2 - My sample
Lane 3 - Shane's sample
Lane 4 - Mihir's sample
Week 5 - 9/24/2012
100112 - Akhi - ok good. Does your Midiprep sequence line up with anything? Is it the right plasmid? I think your primer design looks good. We will order those. For you PCR gel - be sure to add that it is PCR of pGBR22 with M13for/M13rev primers. Also try to crop your gel images better. -- Dr. B
Sequencing Results from Midi Prep:
NNNNNNNNNNNNNNNNNNNNNCTNNGTGGTGGNGGNGGTGGTGCTCGAGTGCGGCCGCAAGCTTGTCGACGGAGCTCGNNNNNNNNNNNGNNNCCACCTTTACTGGAGACCGTCAATGCCAATANNNTNNNGGCATTTTCTTTTGCGTTTTTATTTGTTAACTGTTAATTGTCCTTGTTCAAGGATGCTGTCTTTGACAACAGATGTTTTCTTGCCTTTGATGTTCAGCAGGAAGCTAGGCGCAAACGTTGATTGTTTGTCTGCGTAGAATCCTCTGTTTGTCATATAGCTTGTAATCACGACATTGTTTCCTTTCGCTTGAGGTACAGCGAAGTGTGAGTAAGTAAAGGTTACATCGTTAGGATCAAGATCCATTTTTAACACAAGGCCAGTTTTGTTCAGCGGCTTGTATGGGCCAGTTAAAGAATTAGAAACATAACCAAGCATGTAAATATCGTTAGACGTAATGCCGTCAATCGTCATTTTTGATCCGCGGGAGTCAGTGAACAGGTACCATTTGCCGTTCATTTTAAAGACGTTCGCGCGTTCAATTTCATCTGTTACTGTGTTAGATGCAATCAGCGGTTTCATCACTTTTTTCAGTGTGTAATCATCGTTTAGCTCAATCATACCGAGAGCGCCGTTTGCTAACTCAGCCGTGCGTTTTTTATCGCTTTGCAGAAGTTTTTGACTTTCTTGACGGAAGAATGATGTGCTTTTGCCATAGTATGCTTTGTTAAATAAAGATTCTTCGCCTTGGTAGCCATCTTCAGTTCCAGTGTTTGCTTCAAATACTAAGTATTTGTGGNCTTTATCTTCTACGTAGTGAGGATCTCTCAGCGTATGGGTTGTCGCCTGAGCTGTAGTNGCCNTTCATCGATGAACTGCTGTACATTTTGATACGTTTTTCCGTCACCGTCAAAGATTGATTTATAATCCTCTACACCNNTGATGTTCAAAGAGCTGTCTGATGCTGATACGTTAANNTGNGCAGTGNCAGTGTTNGNTTNNCGNAANGTTTACCGGANAAATCAGTGNANANNAAACGGATTTTTCCGTCANANGNNNTGNGCNTNAACCNNACCNNTNCTTGNNTTNNGNNNTTTTNAGGATANNNTCATTTNCAATCGAAATTTNNCNCNNNNCTTTAAANNNNNNNNNNNNNTTTTTTNCNNNNNNNNNNANNAANTTNNNNNCCNANTTTTTGNNANNAANNNNNAANNCNNNNNNNCNNNNNNNNTTTNNNNTNNNNNNNNNNNNAANNNNAANNNNGNNNNNNNNNNNTTNNNNNNNNNNNNNCCNNNNNNNNNNTTNTNNANNNNNGNCN
It is the right plasmid.
Design Primer Protocl Results: Also posted on google docs.
PCR 1
1st Lane - DNA 1 kb ladder
2nd Lane - .3ng
3rd Lane - 3ng
4th Lane - 30ng
5th Lane - negative control (i might have accidentally added some plasmid)
Week 4 - 9/16/2012
Akhi - ok concentration on midiprep is good. Depending on how much you use for cloning, you may end up needing to make more? -- Dr. B
Did midi prep and have currently sent the obtained plasmid to sequencing. Will post results when available. In the meantime, nano drop reveals
Concentration 53.6ng/uL
Also did the PCR protocol for pLIC sequencing vectors of pNIC-Bsa4. Currently let the samples run through the thermal program and stored in the -20 freezer, next step is to run it with the agarose gel.
Problems occurred. Seems that the machine was programmed wrong, 5min heat for 30 cycles instead of 1 min for 30 cycles. Will see to it on Monday.
Week 3 - 9/10/2012
Akhi - ok good. Include analysis of your RE digest results. Do your pLIC results match up to what they should be? -- Dr. B 091812
Data pending (talk about transformatio
Did a transformation pNIC- Bsa4, left the conicals in the -20 freezer.
RE Digest Results:
9/13/2012
Akhilesh Padhye
RE Digest pGBR22
Lane 1 skip
Lane 2 1kb DNA ladder NEB
Lane 3 1kb DNA ladder NEB
Lane 4 Uncut Plasmid
Lane 5 EcoRI
Lane 6 PvuII
Lane 7 EcoRI + PvuII
Questions for RE Digest: Why was the 3rd band a bid higher even though there was no reduction in size? The uncut plasmid curled up during the process and was able to navigate more quickly, compared to the stretched out EcoRI.
Why did the last lane have a fading band in the end? Possibly due to that sequence being a bit lighter than the rest of the bands.
Did pLIC primer dilution first.
Sequencing Results for
The pLIC reverse sequence did indeed match with what was expected. Blast results yielded a 99.8% identity.
Wrap up of the purification and transformation protocol. Started the enzyme assays. One of which failed, consequently tried again.
Characterization results indicate weight of about 47kDa.
This is the figure from the successful enzyme assay.
This is the data results from the unsucessful enzyme assay.
Week 14 - 11/26/2012
Worked a bit on the surrogate procedure. Predominately spent time on the virtual screening of the target and creating a swiss model. Ligand binding continues to prove unsucessful. Will recheck gold coordinates next week.
Figure 1: Homology model achieved using the SWISS protocol.
More data pending (left in the VDS class laptop).
Week 13 - 11/19/2012
112612 - Good - show some 'data' or image. Dr B
Worked a bit on the latter portion of the surrogate protocol. No other work due to Thanksgiving Break. Data images pending.
Week 12 - 11/12/2012
Akhi, ok - would like to see graph or data from virtual here. -- Dr. B 11/19/12
Moved onto the surrogate protocol. Did the day one and day two steps.
mL required for 0.1 OD reading - 40mL.
(More specific data impending)
1.5 hours required to achieve .346 OD reading.
Weight of Bacteria - 1.5g , 1.19g, 1.16g.
Week 11 - 11/5/2012
Started the cloning and transformation protocol. Then decided it would be more efficient if Shane took care of most of it and I made more backup PCR products instead. First I though I found some leftover PCR sqared, so I tested it, however it was not PCR product at all (picture below confirms it) . Then I used PCR secondary and started the to create PCR squared. However, Shane tested it and found out that the PCR did not work.
Results indicate anonymous samples were not PCR squared. work.
Week 10 - 10/29/2012
PCR squared was finally a success. Two concentrations were achieved.
PCR cleanup concentration 1: Gained through using 100uL of PCR product during cleanup.
PCR cleanup concentration 2: gained through using another 100uL of PCR product during cleanup.
Both concentrations are acceptable, though the 56.4 ng/uL is preferable. My group plans to utilize this latter.
Week 9 - 10/22/2012
Did two PCR cleanups. Unfortunately, both results yielded 0 ng as a concentration. This would mean that either I am missing some important component in my PCR protocols, or I am getting very unlucky. Will try another PCR cleanup next week.
Week 8 - 10/15/2012
Predominately dedicated time to Virtual Drug Screening Refresher. Results are posted on my Google docs. Was going to start PCR cleanup on Saturday but fell ill.
Week 7 - 10/8/2012
101612 - Akhi - ok good job. Good luck with PCR cleanup step and cloning. -- Dr. B
The final four lanes are my PCR^2. The results correlate with the results from PCR secondary and also match up with the predicted length of my gene sequence.
I also worked on virtual drug screening and finished part 1. Currently waiting for results.
Week 6 - 10/1/2012
100912 - Akhi, ok great. I am glad you guys got this to work. Crop your gels a little more so the gel is more visible. When you guys move on to cloning - use the NEW T4 DNA Poly and the new dGTP and new dCTP.
- Dr. B
Ran primary PCR and got these results.
PCR Primary Results
Lane 1 - 100bp ladder
Lane 2 - Shane's sample
Lane 3 - Mihir's sample
Lane 4 - My sample (not even visible), perhaps attributed to the low temperature I used during the pcr program.
Secondary PCR (done using Mihir's stock)
Lane 1 - 100bp ladder
Lane 2 - My sample
Lane 3 - Shane's sample
Lane 4 - Mihir's sample
Week 5 - 9/24/2012
100112 - Akhi - ok good. Does your Midiprep sequence line up with anything? Is it the right plasmid? I think your primer design looks good. We will order those. For you PCR gel - be sure to add that it is PCR of pGBR22 with M13for/M13rev primers. Also try to crop your gel images better. -- Dr. B
Sequencing Results from Midi Prep:
NNNNNNNNNNNNNNNNNNNNNCTNNGTGGTGGNGGNGGTGGTGCTCGAGTGCGGCCGCAAGCTTGTCGACGGAGCTCGNNNNNNNNNNNGNNNCCACCTTTACTGGAGACCGTCAATGCCAATANNNTNNNGGCATTTTCTTTTGCGTTTTTATTTGTTAACTGTTAATTGTCCTTGTTCAAGGATGCTGTCTTTGACAACAGATGTTTTCTTGCCTTTGATGTTCAGCAGGAAGCTAGGCGCAAACGTTGATTGTTTGTCTGCGTAGAATCCTCTGTTTGTCATATAGCTTGTAATCACGACATTGTTTCCTTTCGCTTGAGGTACAGCGAAGTGTGAGTAAGTAAAGGTTACATCGTTAGGATCAAGATCCATTTTTAACACAAGGCCAGTTTTGTTCAGCGGCTTGTATGGGCCAGTTAAAGAATTAGAAACATAACCAAGCATGTAAATATCGTTAGACGTAATGCCGTCAATCGTCATTTTTGATCCGCGGGAGTCAGTGAACAGGTACCATTTGCCGTTCATTTTAAAGACGTTCGCGCGTTCAATTTCATCTGTTACTGTGTTAGATGCAATCAGCGGTTTCATCACTTTTTTCAGTGTGTAATCATCGTTTAGCTCAATCATACCGAGAGCGCCGTTTGCTAACTCAGCCGTGCGTTTTTTATCGCTTTGCAGAAGTTTTTGACTTTCTTGACGGAAGAATGATGTGCTTTTGCCATAGTATGCTTTGTTAAATAAAGATTCTTCGCCTTGGTAGCCATCTTCAGTTCCAGTGTTTGCTTCAAATACTAAGTATTTGTGGNCTTTATCTTCTACGTAGTGAGGATCTCTCAGCGTATGGGTTGTCGCCTGAGCTGTAGTNGCCNTTCATCGATGAACTGCTGTACATTTTGATACGTTTTTCCGTCACCGTCAAAGATTGATTTATAATCCTCTACACCNNTGATGTTCAAAGAGCTGTCTGATGCTGATACGTTAANNTGNGCAGTGNCAGTGTTNGNTTNNCGNAANGTTTACCGGANAAATCAGTGNANANNAAACGGATTTTTCCGTCANANGNNNTGNGCNTNAACCNNACCNNTNCTTGNNTTNNGNNNTTTTNAGGATANNNTCATTTNCAATCGAAATTTNNCNCNNNNCTTTAAANNNNNNNNNNNNNTTTTTTNCNNNNNNNNNNANNAANTTNNNNNCCNANTTTTTGNNANNAANNNNNAANNCNNNNNNNCNNNNNNNNTTTNNNNTNNNNNNNNNNNNAANNNNAANNNNGNNNNNNNNNNNTTNNNNNNNNNNNNNCCNNNNNNNNNNTTNTNNANNNNNGNCN
It is the right plasmid.
Design Primer Protocl Results:
Also posted on google docs.
Upstream: TACTTCCAATCCATGAAAAACTGGGTTAAAG
Forward:
Mg++ 0
Downstream: TATCCACCTTTACTGTTAGTACAGGTATGCTTTT
Reverse Complement of the reverse primer: AAAAGCATACCTGTACTAACAGTAAAGGTGGATA
Mg++ 0
PCR 1
1st Lane - DNA 1 kb ladder
2nd Lane - .3ng
3rd Lane - 3ng
4th Lane - 30ng
5th Lane - negative control (i might have accidentally added some plasmid)
Week 4 - 9/16/2012
Akhi - ok concentration on midiprep is good. Depending on how much you use for cloning, you may end up needing to make more? -- Dr. B
Did midi prep and have currently sent the obtained plasmid to sequencing. Will post results when available. In the meantime, nano drop reveals
Concentration 53.6ng/uL
Also did the PCR protocol for pLIC sequencing vectors of pNIC-Bsa4. Currently let the samples run through the thermal program and stored in the -20 freezer, next step is to run it with the agarose gel.
Problems occurred. Seems that the machine was programmed wrong, 5min heat for 30 cycles instead of 1 min for 30 cycles. Will see to it on Monday.
Week 3 - 9/10/2012
Akhi - ok good. Include analysis of your RE digest results. Do your pLIC results match up to what they should be? -- Dr. B 091812
Data pending (talk about transformatio
Did a transformation pNIC- Bsa4, left the conicals in the -20 freezer.
RE Digest Results:
9/13/2012
Akhilesh Padhye
RE Digest pGBR22
Lane 1 skip
Lane 2 1kb DNA ladder NEB
Lane 3 1kb DNA ladder NEB
Lane 4 Uncut Plasmid
Lane 5 EcoRI
Lane 6 PvuII
Lane 7 EcoRI + PvuII
Questions for RE Digest: Why was the 3rd band a bid higher even though there was no reduction in size? The uncut plasmid curled up during the process and was able to navigate more quickly, compared to the stretched out EcoRI.
Why did the last lane have a fading band in the end? Possibly due to that sequence being a bit lighter than the rest of the bands.
Did pLIC primer dilution first.
Sequencing Results for
The pLIC reverse sequence did indeed match with what was expected. Blast results yielded a 99.8% identity.
LICrAAP-pLIC-rev:
NNNNNNNTGNNNNNTTTAGNNGAGATATACATATGCACCATCATCATCATCATTCTTCTGGTGTAGATCTGGGTACCGAG
AACCTGTACTTCCAATCCATGGAGACCGACGTCCACATATACCTGCCGTTCACTATTATTTAGTGAAATGAGATATTATGA
TATTTTCTGAATTGTGATTAAAAAGGCAACTTTATGCCCATGCAACAGAAACTATAAAAAATACAGAGAATGAAAAGAAA
CAGATAGATTTTTTAGTTCTTTAGGCCCGTAGTCTGCAAATCCTTTTATGATTTTCTATCAAACAAAAGAGGAAAATAGACC
AGTTGCAATCCAAACGAGAGTCTAATAGAATGAGGTCGAAAAGTAAATCGCGCGGGTTTGTTACTGATAAAGCAGGCAA
GACCTAAAATGTGTAAAGGGCAAAGTGTATACTTTGGCGTCACCCCTTACATATTTTAGGTCTTTTTTTATTGTGCGTAACTA
ACTTGCCATCTTCAAACAGGAGGGCTGGAAGAAGCAGACCGCTAACACAGTACATAAAAAAGGAGACATGAACGATGA
ACATCAAAAAGTTTGCAAAACAAGCAACAGTATTAACCTTTACTACCGCACTGCTGGCAGGAGGCGCAACTCAAGCGTTT
GCGAAAGAAACGAACCAAAAGCCATATAAGGAAACATACGGCATTTCCCATATTACACGCCATGATATGCTGCNNTCCCTG
AACAGCAAAAAAATGAAAAATATAAAGTTCCTGAGTTCGATTCGTCCACAATTAAAAATATCTCTTCTGCAAAAGGCCTGGA
CGTTTGGGACAGCTGGCCATTACAAAACACTGACGGCACTGTCGCAAACTATCACGGCTACCACATCGTCTTTGCATTAGC
CGGAGATCCTAAAAATGCGGATGANNCATCGATTTACATGTTCTATCAAAAAGTCGGCGAAACTTCTATTGACAGCTGGAAA
ACGCTGGCNGCGTCTTTNANGACAGCGACAANTCGATGCAATGATNCTATCCTAAANACCAANCNCAANNATGNCNNNNAN
CNCNTTTNCNTCTGANGAAAANCCNTNNTCTANNCTGATNNNCNNANNNTNNGNANNNANNNCTGANANTGNNNANNANNN
ATCANCNTNNNNNNNNNNNNANNNNNNNGNNNANNGNNNNNNAANNNNNNTNNNNGGNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNGNNNTNNNNNNNGGNNNNNNNNNNCNNNNNNNNNNNNNNNNNNN
LICfAAP-pLIC-for
NNNNNNNNNNNNGGNNCTCNGTGGTGGTGGTGGTGGTGCTCGAGTGCGGCCGCAAGCTTGTCGACGGAGCTCGAATTCG
GATCCGTATCCACCTTTACTGGAGACCGTCAATGCCAATAGGATATCGGCATTTTCTTTTGCGTTTTTATTTGTTAACTGTTAATT
GTCCTTGTTCAAGGATGCTGTCTTTGACAACAGATGTTTTCTTGCCTTTGATGTTCAGCAGGAAGCTAGGCGCAAACGTTGATT
GTTTGTCTGCGTAGAATCCTCTGTTTGTCATATAGCTTGTAATCACGACATTGTTTCCTTTCGCTTGAGGTACAGCGAAGTGTGA
GTAAGTAAAGGTTACATCGTTAGGATCAAGATCCATTTTTAACACAAGGCCAGTTTTGTTCAGCGGCTTGTATGGGCCAGTTAA
AGAATTAGAAACATAACCAAGCATGTAAATATCGTTAGACGTAATGCCGTCAATCGTCATTTTTGATCCGCGGGAGTCAGTGAA
CAGGTACCATTTGCCGTTCATTTTAAAGACGTTCGCGCGTTCAATTTCATCTGTTACTGTGTTAGATGCAATCAGCGGTTTCATCA
CTTTTTTCAGTGTGTAATCATCGTTTAGCTCAATCATACCGAGAGCGCCGTTTGCTAACTCAGCCGTGCGTTTTTTATCGCTTTGCA
GAAGTTTTTGACTTTCTTGACGGAAGAATGATGTGCTTTTGCCATAGTATGCTTTGTTAAATAAAGATTCTTCGCCTTGGTAGCCAT
CTTCAGTTCCAGTGTTTGCTTCAAATACTAAGTATTTGTGGCCTTTATCTTCTACGTAGTGAGGATCTCTCAGCGTATGGTTGTCGC
CTGAGCTGTAGTTGCCTTCATCGATGAACTGCTGTACATTTTGATACGTTTTTCCGTCACCGTCAAANNATTGATTTNNNATCCTCT
ACACCGTTGATGTTCAAAGAGCTGTCTGANGCTGATACGTTAACTTNTGCAGTNTCAGTGNTTGNTTGNCGTANGNTTACCGNA
NNAATCANTGNNNAATNAACGNATTTTNCGTCNNATGTAAATGNGNTGANNTGANNNTTNNNNNNTNNNNNTTNNNNNNNNCAT
TNGCATCNNNNGTCNNNNNNNTTNNNNNNNGNNNNNNTNNNNCNNNNNNNNANNNNNCCNANTTTNNNNNNNNNNNNNNNNA
NNNNNNNCNNNNNNNNNNNNNNNGNANNNNNNNNNCNNNNNNNNNNNNNNNNNCNNNNN
Week 2 - 9/3/2012
Not much lab work - only diluted primers and got them ready for primer DNA sequencing.
Analyzed a DNA sequence in the computer lab.
Week 1 - 8/27/2012
Basic Info:
Target (protein/gene name): Protein Tyrosine Phosphatase - lmo1800
*NCBI Gene # or RefSeq#: Project:61583, NC_003210.1, Gene ID: 985934
*Protein ID (NP or XP #) or Wolbachia#: NP_465325.1
*Organism: Listeria monocytogenes
*EC#: 3.1.3.48
*Amino Acid Sequence:
>gi|16803840|ref|NP_465325.1| hypothetical protein lmo1800 [Listeria monocytogenes EGD-e]
MKNWVKVTGAGVLSATLLLGGCGAQSEEKAEANVKTEQTLKPGSQIKLEGAVNVRDLGGYKTTDGLTIKPHKLIRSAELANLSDSDKKKLVNTYDLSHIVDFRTSSEVATKPDPKLTDVDYTHDSVMKDNGTSTSTQDLTASLAKMDNPETFLINANKSFITDETSIQAYKDFFDILLANQDGSVLWHCTAGKDRAGFGTALVLSALGVDKNTVIDDYMLSNKYRADENKKAIEAVAAKTDNKKVIDGMTAVMEVRESYINAAFDEINAKYGSMDNFLKEKLGLTDAKKEQLKKAYLY
*CDS Gene Sequence:
Partial
>gi|16802048:c1873620-1872724 Listeria monocytogenes EGD-e, complete genomeATGAAAAATTGGGTAAAAGTAACAGGAGCAGGGGTACTAAGCGCGACTTTACTATTAGGTGGATGCGGGG CGCAGTCAGAAGAAAAAGCAGAAGCAAATGTTAAAACCGAGCAAACACTCAAACCAGGAAGCCAAATTAA ATTAGAAGGCGCTGTAAATGTCCGGGACTTAGGCGGATACAAAACAACGGATGGACTAACCATTAAGCCA CATAAACTCATTAGAAGTGCCGAACTCGCTAACTTAAGTGATTCGGATAAAAAGAAACTCGTAAATACAT ATGATTTGTCTCATATAGTTGATTTCCGAACAAGTTCAGAAGTCGCAACGAAACCAGATCCGAAACTCAC AGATGTAGACTATACGCACGATTCTGTGATGAAAGATAATGGAACATCTACAAGCACACAAGATTTAACT GCTAGCCTAGCCAAAATGGATAATCCAGAGACATTTCTCATTAATGCTAATAAGAGCTTTATTACAGATG AAACTTCGATACAAGCCTATAAAGATTTTTTTGATATACTACTAGCAAACCAAGATGGTTCTGTTCTTTG GCACTGTACAGCTGGAAAAGACCGAGCTGGATTTGGAACCGCCCTCGTTCTTTCAGCGTTAGGTGTGGAT AAAAACACGGTCATTGACGATTATATGCTGTCCAATAAATATCGTGCTGACGAAAATAAAAAAGCAATTG AAGCCGTTGCAGCAAAAACAGATAATAAAAAAGTGATTGATGGAATGACAGCCGTAATGGAAGTTCGTGA ATCTTATATCAATGCAGCCTTCGATGAAATTAATGCAAAATATGGTTCGATGGACAACTTTTTAAAAGAA AAACTCGGACTAACCGATGCTAAAAAAGAACAACTAAAAAAAGCATATCTTTATTAA
Full Info:
http://vdsstream.wikispaces.com/Target+-+Protein+Tyrosine+Phosphatase+%28Listeria+monocytogenes%29